• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.95-95
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• 1995

뇌신경 변성 / 퇴행과 관련된 중요한 병인론 중의 하나는 변성 과정에서 형성된 유리기(free radical)로 인한 항산화계의 평형 소실로 알려져 있다. Aspalatone (APT)의 예상되는 항산화 효능을 검정하기 위하여 본 실험에서는 Kainic acid (KA) 유발 뇌변성 모델을 적용하였다. KA 모델은 변연계의 간질성 경련과 신경세포 변성에 대하여 재현성 있는 병변 모델을 제공해 주며, 이와 같은 신경세포의 병독 기전에 산소 유리기가 관여함이 강력히 시사되고 있기 때문이다. KA 투여로 인하여 지속적이고도 전형적인 간질성 경련이 관찰되고 1일 이내에 높은 치사율을 보였으나 APT으로 인하여 그 간질성 경련 행위와 비율이 억제되고 KA 유발 치사율도 억제되었다. 최종 KA 투여 3일 후에 얻어진 흰쥐 해마 및 대뇌 피질에서 항산화 효소인 Superoxide dismutase (SOD), Catalase (Cat.), Glutathione peroxidase (GSH-PX) 및 과산화지질의 지표인 Malondialdehyde (MDA)를 검정하였다. 대조군에 비하여 KA는 뇌조직의 SOD-1을 유도하였으나, Cat.와 GSH-PX의 활성은 현저히 유도되지 않았고, 반면에 MDA 치는 현저히 증가하였다. 즉, Cat., GSH-PX와 같은 $H_2O$$_2$중화제가 동반 유도하지 않는 SOD의 유도는 세포내 축적되는 $H_2O$$_2$로 인하여 Fenton/Haber-Weiss 반응을 가속화하여 과산화지질화를 촉진함을 시사한다. APT 병용 투여로 SOD는 현저히 유도되지 않았으나 특히 Cat.가 현저히 유도되어지고 MDA는 억제되었다. 이와 같은 생화학적인 결과는 다음의 형태학적인 소견과 일치한다. Fos 관련 항원 (FRA)와 SOD-1을 면역세포화학 (Immunocytochemistry)적 방법으로 이중 표식 (double-labelling) 하였다. FRA는 KA로 인한 신경세포의 자극에 대한 지표로 응용하였고, SOD-1은 퇴행성 뇌질환에서 산화적 손상의 지표로 사용하였다. KA 투여로 해마의 dentate gyrus (DG) 내에 강한 면역환성 (immunoreactivity)이 나타났고 pyramidal cell layer (PCL)와 glia에 SOD-1이 강하게 염색되었다. APT 병용 투여로 상당수의 경련이 일어나지 않은 흰쥐는 해마의 DG에 FRA가 경미하게 염색되었고, PCL에 SOD-1도 경미하게 나타났으나, 경련이 나타난 쥐에서는 KA만을 투여한 흰쥐와 구별되지 않았다. 이상의 APT의 항산화 효과는 KA로 인한 뇌세포 변성 개선에 중요한 인자로 작용할 것으로 사료되나, 보다 명확한 APT의 기전을 검색하고 직접 임상에 응응하기 위하여는 보다 다양한 실험 조건이 보완되어야 찰 것으로 생각된다.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.6
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• pp.483-491
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• 2019

Cordycepin exerts neuroprotective effects against excitotoxic neuronal death. However, its direct electrophysiological evidence in Alzheimer's disease (AD) remains unclear. This study aimed to explore the electrophysiological mechanisms underlying the protective effect of cordycepin against the excitotoxic neuronal insult in AD using whole-cell patch clamp techniques. ${\beta}$-Amyloid ($A{\beta}$) and ibotenic acid (IBO)-induced injury model in cultured hippocampal neurons was used for the purpose. The results revealed that cordycepin significantly delayed $A{\beta}$ + IBO-induced excessive neuronal membrane depolarization. It increased the onset time/latency, extended the duration, and reduced the slope in both slow and rapid depolarization. Additionally, cordycepin reversed the neuronal hyperactivity in $A{\beta}$ + IBO-induced evoked action potential (AP) firing, including increase in repetitive firing frequency, shortening of evoked AP latency, decrease in the amplitude of fast afterhyperpolarization, and increase in membrane depolarization. Further, the suppressive effect of cordycepin against $A{\beta}$ + IBO-induced excessive neuronal membrane depolarization and neuronal hyperactivity was blocked by DPCPX (8-cyclopentyl-1,3-dipropylxanthine, an adenosine $A_1$ receptor-specific blocker). Collectively, these results revealed the suppressive effect of cordycepin against the $A{\beta}$ + IBO-induced excitotoxic neuronal insult by attenuating excessive neuronal activity and membrane depolarization, and the mechanism through the activation of $A_1R$ is strongly recommended, thus highlighting the therapeutic potential of cordycepin in AD.

• Bulletin of the Korean Chemical Society
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• v.10 no.2
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• pp.191-196
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• 1989

We obtained the new complexes, $Fe{\eta}^4-R,R'-TPSCp](CO)_3$(R,R'-TPSCp = 1,1-disubstituted 2,3,4,5-Tetraphenyl-1-Silacyclopenta-2,4-diene; R = Ph, R' = Cl, R = R' = Cl) from the reaction of the corresponding R,R'-TPSCp with ironpentacarbonyl under reflux in toluene. Also, the analogous complexes with R = R' = Me and R = Me, R' = Cl were obtained in an identical manner. We have determined the crystal structure of $Fe[Ph(Cl)-TPSCp](CO_)3$ by using Mo ka, ${\lambda}$ = 0.71069${\AA}$, where the unit cell was found to be monoclinic with a = 9.042 (6)${\AA}$, b = 19.870 (9)${\AA}$, c = 17.426 (9)${\AA}$ and ${\beta}$ = 96.28(4)$^{\circ}$. The butadiene moiety of TPSCp ring is planar and the dihedral angle of the butadiene plane and C4-Si-C25 plane was opened up to 41.8$^{\circ}$. The C-C distances in the butadiene moiety were found to be 1.4346, 1.462, and 1.440 ${\AA}$, respectively. It may be said that the four ${\pi}$-electrons are delocalized over the four carbons in five membered ring through coordination with ironcarbonyl. In this complex Fe is either in distorted tetrahedron environment with the centroid of the four C-atom butadiene moiety and three carbons of the three carbonyls or in distorted square-pyramidal environment with two midpoints of double bonds of the butadiene moiety and two carbons of carbonyl defining the base of the pyramid and the carbon of remaining carbonyl the apex.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1112-1117
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• 2008

Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

• Biomedical Science Letters
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• v.6 no.1
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• pp.11-17
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• 2000

SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼991 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-21.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.130-130
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• 1995

AM, SM 및 ASA는 수용액중에서 겉보기 1차반응에 따라 분해되었으며 보존온도가 높을수록 분해가 촉진되는 온도 의존성을 나타내었다. AM의 분해경로는 pH 1.22 및 pH 7.0 이상에서는 AM$\longrightarrow$ SM $\longrightarrow$ SA의 경로로 주로 분해되었으며 pH 2.01 - 6.08의 범위에서는 AM $\longrightarrow$ASA$\longrightarrow$SA의 경로로 분해되는 양상을 보였다. 또 pH가 분해에 미치는 영향을 pH-rate profile로 나타낸 결과 AM, SM 및 ASA의 최대안정 pH는 각각 4.0, 3.0, 2.0 부근이 있고 이 조건에서의 분해 반감기는 114, 168, 113 hr로 나타났다. 전체적으로 보면 pH 2.0 이하에서는 ASA가 AM 보다 약간 안정한 편이나 pH 2.0-8.0 사이에서는 AM의 분해속도가 ASA보다 현저히 낮았다. 또 AM은 pH 7.0 이상에서, SM은 pH 6.0 이상에서, ASA는 9.0 이상에서 특수염기촉매반응에 따라 분해가 이루어지는 것을 알 수 있었다. 이온강도($\mu$)의 영향으로는 pH 7.0에서 이온강도가 0.115에서 1.0으로 증가할수록 $\mu$$^{1}$2/에 대해 AM의 분해속도정수가 직선적으로 완만하게 감소되었다. 또 완충수용액 중 AM의 가수분해 억제효과를 검토하기 위해 시클로덱스트린류를 첨가하였을 때, $\beta$-시클로덱스트린과 히드록시프로필기-$\beta$-시클로덱스트린은 AM의 분해를 각각 1.6배 및 1.1배 촉진시켜 촉매적으로 작용하였으며 디메칠-$\beta$-시클로덱스트린은 약 3.2배 분해속도를 억제시켜 안정화제로 작용하였다.Zn^{2+}$, soybean trypsin inhibtor에 의해 25～50% 정도, serine proteinase inhibitor인 phenylmethylsulfonyl floride에 의해 80%정도 활성이 억제되는 특성이 있음을 규명하였다.면역환성 (immunoreactivity)이 나타났고 pyramidal cell layer (PCL)와 glia에 SOD-1이 강하게 염색되었다. APT 병용 투여로 상당수의 경련이 일어나지 않은 흰쥐는 해마의 DG에 FRA가 경미하게 염색되었고, PCL에 SOD-1도 경미하게 나타났으나, 경련이 나타난 쥐에서는 KA만을 투여한 흰쥐와 구별되지 않았다. 이상의 APT의 항산화 효과는 KA로 인한 뇌세포 변성 개선에 중요한 인자로 작용할 것으로 사료되나, 보다 명확한 APT의 기전을 검색하고 직접 임상에 응응하기 위하여는 보다 다양한 실험 조건이 보완되어야 찰 것으로 생각된다. 항우울약들의 항혈소판작용은 PKC-기질인 41-43 kD와 20 kD의 인산화를 억제함에 기인되는 것으로 사료된다.다. 것으로 사료된다.다.바와 같이 MCl에서 작은 Dv 값을 갖는데, 이것은 CdCl$_{4}$$^{2-}$ 착이온을 형성하거나 ZnCl$_{4}$$^{2-}$ , ZnCl$_{3}$$^{-}$같은 이온과 MgCl$^{+}$, MgCl$_{2}$같은 이온종을 형성하기 때문인것 같다. 한편 어떠한 용리액에서던지 NH$_{4}$$^{+}$의 경우 Dv값이 제일 작았다. 바. 본 연구의 목적중의 하나인 인체유해 중금속이온인 Hg(II), Cd(II)등이 NaCl같은 염화물이 함유된 시료용액에 공해이온으로 존재할 경우 흡착에 의한 제거가 가능하다. 한편 이같

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.305-311
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• 2007

Glucocorticoids (GCs) alter metabolism, synaptogenesis, apoptosis, neurogenesis, and dendritic morphology in the hippocampus. To better understand how glucocorticoids regulate these aspects of hippocampal biology, we studied gene expression patterns in the CA3 (Hippocampal pyramidal cell field CA3) and dentate gyrus (DG). Litter-matched Lewis inbred rats treated for 20 days with either 9.5 mg per day sustained-release corticosterone or placebo pellets were compared with high-density oligonucleotide microarray analysis (Rat Neurobiology U34 Arrays, Affymetrix). In placebo-treated rats, 32 genes were expressed at greater levels in CA3 than DG, whereas 3 genes were expressed at great levels in DC than CA3. Regional differences were also apparent in corticosterone-induced changes in the hippocampal transcriptome. Six genes in CA3 and 41 genes in DC were differentially regulated by corticosterone. As per the glucocorticoid effects on gene transcription in the brain, forty three of these genes were upregulated, and 4 genes were downregulated. Genes differentially expressed in hippocampus included those for 13 neurotransmitter proteins, 5 ion channel related proteins, 4 transcription factors, 3 neurotrophic factors, 1 cytokine, 1 apoptosis related protein, and 5 genes involved in synaptogenesis. Interestingly, GCs can have suppressive effects on brain BDNF mRNA transcription, one of the neurotrophic factors. These results indicate the diversity of targets affected by chronic exposure to corticosterone and highlight important regional differences in hippocampal neurobiology.

• Journal of Ginseng Research
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• v.28 no.2
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• pp.94-103
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• 2004

Korean red ginseng is known to have anti-stress and memory enhancing effects. Recent studies suggested that stress-induced inhibition of adult neurogenesis in hippocampus may contribute, in part, to decreased negative feedback inhibition of HPA axis. In order to elucidate the mechanism of Korean red ginseng in anti-stress and memory enhancing effects, we observed the effects of repeated treatment of Korean red ginseng total saponin (GTS, 50 mg/kg, i.p.) in response to repeated unpredictable stress for 10 days. Male Sprague-Dawley rats (230 - 260 g) received with either GTS (50 mg/kg, i.p.) or vehicle (1 ml/kg, i.p.) 1 h before stress for 10 days. Rats were injected with bromodeoxyuridine (BrdU, 50 mg/kg, i.p.) 16-18 he after last stress procedure, and were sacrificed 2 hr later by perfusion. Immunohistochemistry of BrdU was done to measure proliferation of neural progenitor cells in hippocampus, which was used as an index of neurogenesis. Repeated GTS treatment for 10 days increased neurogenesis in subgranular zone area of dentate gyrus (SGZ), but not hilus, compared with vehicle-treated rats. Repeated unpredictable stress did not affect the neurogenesis compared with controls, while repeated GTS treatment increased neurogenesis in SGZ in repeated unpredictable stress-exposed group. BDNF mRNA was also measured in subregions of hippocampus by in situ hybridization. BDNF mRNA expression in CA3 and CA1 pyramidal cell layer was increased by repeated GTS treatment but not in dentate granule cell layer. Repeated unpredictable stresses significantly decreased BDNF mRNA expression in all subregions of hippocampus, but repeated GTS treatment did not prevent stress-induced BDNF mRNA downregulation. Given that repeated GTS treatment increased proliferation of neural progenitor cells in repeated unpredictable stress-exposed rats in the presence of decreased BDNF mRNA expression in dentate granule cell layer, it raise the possibility that BDNF may not playa significant role in GTS-mediated increase of neurogenesis in adult rat hippocampus. Also, these results suggest that repeated GTS treatment increased neurogenesis of SGZ and BDNF mRNA expression, which may account for memory enhancing effect of Korean red ginseng. In addition, repeated GTS treatment appears not to have anti-stress effects in terms of neurotrophin, but GTS-mediated increase of neurogenesis in hippocampus may contribute to increase negative feedback inhibition of HPA axis.

• The Korean Journal of Zoology
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• v.31 no.2
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• pp.111-121
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• 1988

This study was attempted to investigate the regional distribution and shapes of pancreatic endocrifle cells in the hedgehog, Erinaceus koneanus by the immunocytochemical PAP methods (Nalane, 1968; Stemberger, 1979). The tissue specimen taken from the splenic and duodenal regions of pancreas(proximai, middle and distal portions, respectively) were fixed with Bouin solution and the sections(5$\mu$ m) were followed by simple and double staining with 2 substrates, DAB and 4-CI-1-naphthol. The results were as following: Glucagon(A) cells, 13 $\mu$ m x 9.5 $\mu$ m in size, were found in the islets periphery and among the exocrine parenchyma. A cells were abundant in all the portions of splenic region and distal portion of duodenal region in contrast to a few in the proximal and middle portion of duodenal region. The shapes of the A cells were round, oval and pyramidal types. Insulin(B) cells, 11.6$\mu$m x 9.4$\mu$m in sise, were round or oval in shape and located throughout the islets. B cells were the most numerous cell types in all portions of splenic region and distal protion of duodenal region as compared with the other portions. Somatostatin(D) cells, 12.6$\mu$m x 9.1 $\mu$m in size, were round or oval in shape and found in the islets periphery and scattered in the exocrine parenchyma. These cells were rare in all the portions of splenic and duodenal region. Pancreatic polypeptide(PP) cells of various type, 12.8$\mu$ m x 8.5 $\mu$ m in size, were found in the islets periphery and among the exocrine parenchyma. PP cells were numerous in the proximal and middle portion of duodenal region, but rarely scattered in the other portions.